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Abstract The object of this study is to prepare antisera to purified glycosphingolipids for immunocytochemical studies. Rabbits were immunized with ganglioside GM1 complexed either to methylated BSA or to a glycoprotein isolated from human erythrocytes, and to asialo GM1 complexed to the erythrocyte glycoprotein. IgG and IgM antibody fractions were separated by chromatography on Sephadex G-200 and their properties were determined by complement fixation. IgG antibodies to asialo GM1 were highly specific and exhibited little cross-reactivity with GM1 or other glycolipids, but IgM antibodies cross-reacted extensively with gangliosides GM1 and GD1b. Complement fixation of IgG anti-asialo GM1 with asialo GM1 was completely inhibited by about 15 nmoles of ganglio-N-tetraose, the carbohydrate moiety of this glycolipid. Both IgG and IgM antibodies to GM1 cross-reacted extensively with asialo GM1 and GD1b, but specific IgG antibodies to GM1 were revealed by absorption with asialo GM1. The specific and cross-reacting IgG antibodies to GM1 were inhibited by the pentaose moiety of this glycoplipid, but these antibodies were much harder to inhibit than IgG anti-asialo GM1. These results demonstrate that it is possible to prepare specific antisera to purified glycolipids with a single administration of antigen, in contrast to previous studies in which repeated injections of ganglioside mixtures were employed.
Naiki et al. (Mon,) studied this question.